Department of Biotechnology, Axis Institute of Higher Education, Kanpur, Uttar Pradesh, India.
Department of Biotechnology, Faculty of Engineering and Technology Rama University, Mandhana, Kanpur, Uttar Pradesh, India.
Prog Mol Biol Transl Sci. 2024;208:83-107. doi: 10.1016/bs.pmbts.2024.07.006. Epub 2024 Aug 21.
Fungi contain a wide range of bioactive secondary metabolites (SMs) that have numerous applications in various fields, including agriculture, medicine, human health, and more. It is common for genes responsible for the production of secondary metabolites (SMs) to form biosynthetic gene clusters (BGCs). The identification and analysis of numerous unexplored gene clusters (BGCs) and their corresponding substances (SMs) has been significantly facilitated by the recent advancements in genomic and genetic technologies. Nevertheless, the exploration of secondary metabolites with commercial value is impeded by a variety of challenges. The emergence of modern CRISPR/Cas technologies has brought about a paradigm shift in fungal genetic engineering, significantly streamlining the process of discovering new bioactive compounds. This study begins with an examination of fungal biosynthetic gene clusters (BGCs) and their interconnections with the secondary metabolites (SMs) they generate. Following that, a brief summary of the conventional methods employed in fungal genetic engineering is provided. This study explores various sophisticated CRISPR/Cas-based methodologies and their utilization in examining the synthesis of secondary metabolites (SMs) in fungi. The chapter provides an in-depth analysis of the limitations and obstacles encountered in CRISPR/Cas-based systems when applied to fungal genetic engineering. It also proposes promising avenues for future research to optimize the efficiency of these systems.
真菌中含有广泛的生物活性次生代谢产物 (SMs)，这些产物在农业、医学、人类健康等各个领域都有多种应用。负责次生代谢产物 (SMs) 产生的基因通常形成生物合成基因簇 (BGCs)。基因组和遗传技术的最新进展极大地促进了对大量未探索的基因簇 (BGCs) 及其相应物质 (SMs) 的识别和分析。然而，具有商业价值的次生代谢产物的探索受到多种挑战的阻碍。现代 CRISPR/Cas 技术的出现彻底改变了真菌遗传工程，大大简化了发现新生物活性化合物的过程。本研究首先考察了真菌生物合成基因簇 (BGCs) 及其与产生的次生代谢产物 (SMs) 之间的相互关系。随后，简要总结了真菌遗传工程中使用的传统方法。本研究探讨了各种复杂的基于 CRISPR/Cas 的方法及其在研究真菌次生代谢产物 (SMs) 合成中的应用。该章节深入分析了基于 CRISPR/Cas 的系统在应用于真菌遗传工程时遇到的局限性和障碍，并提出了优化这些系统效率的未来研究的有前景的途径。
